Subcellular localization and duration of mu-calpain and m-calpain activity after traumatic brain injury in the rat: a casein zymography study.
Casein zymographic assays were performed to identify changes in mu-calpain and m-calpain activity in naive, sham-injured, and injured rat cortex at 15 minutes, 3 hours, 6 hours, and 24 hours after unilateral cortical impact brain injury. Cortical samples ipsilateral and contralateral to the site of injury were separated into cytosolic and total membrane fractions. Marked increases in mu-calpain activity in cytosolic fractions in the ipsilateral cortex occurred as early as 15 minutes, became maximal at 6 hours, and decreased at 24 hours to levels observed at 15 minutes after injury. A similar temporal profile of cytosolic mu-calpain activity in the contralateral cortex was observed, although the increases in the contralateral cortex were substantially lower than those in the ipsilateral cortex. Differences were also noted between cytosolic and total membrane fractions. The detection of a shift in mu-calpain activity to the total membrane fraction first occurred at 3 hours after traumatic brain injury and became maximal at 24 hours after traumatic brain injury. This shift in mu-calpain activity between the two fractions could be due to the redistribution of mu-calpain from the cytosol to the membrane. m-Calpain activity was detected only in cytosolic fractions. m-Calpain activity in cytosolic fractions did not differ significantly between ipsilateral and contralateral cortices, and increased in both cortices from 15 minutes to 6 hours after injury. Relative magnitudes of m-calpain versus mu-calpain activity in cytosolic fractions differed at different time points after injury. These studies suggest that traumatic brain injury can activate both calpain isoforms and that calpain activity is not restricted to sites of focal contusion and cell death at the site of impact injury but may represent a more global response to injury.
Zhao
X
X
Vivian L. Smith Center for Neurologic Research, University of Texas-Houston Health Science Center, Department of Neurosurgery 77030, USA.
Posmantur
R
R
Kampfl
A
A
Liu
S J
SJ
Wang
K K
KK
Newcomb
J K
JK
Pike
B R
BR
Clifton
G L
GL
Hayes
R L
RL
D000818:Animals; D015153:Blotting, Western; D001921:Brain / Q000201:enzymology* / Q000648:ultrastructure*; D001930:Brain Injuries / Q000201:enzymology*; D002154:Calpain / Q000032:analysis / Q000378:metabolism*; D002462:Cell Membrane / Q000201:enzymology; D002540:Cerebral Cortex / Q000201:enzymology / Q000648:ultrastructure; D003600:Cytosol / Q000201:enzymology; D003720:Densitometry; D006863:Hydrogen-Ion Concentration; D007527:Isoenzymes / Q000032:analysis / Q000378:metabolism*; D007700:Kinetics; D008297:Male; D051381:Rats; D017207:Rats, Sprague-Dawley
D007527:Isoenzymes; D002154:Calpain